Download Advanced Time-Correlated Single Photon Counting Applications by Wolfgang Becker PDF

By Wolfgang Becker

This ebook is an try and bridge the distance among the instrumental ideas of multi-dimensional time-correlated unmarried photon counting (TCSPC) and common purposes of the strategy. Written through an originator of the procedure and by way of sucessful clients, it covers the fundamental rules of the procedure, its interplay with optical imaging tools and its program to quite a lot of experimental projects in existence sciences and medical research.

The publication is usually recommended for all clients of time-resolved detection strategies in biology, bio-chemistry, spectroscopy of stay platforms, reside telephone microscopy, scientific imaging, spectroscopy of unmarried molecules, and different purposes that require the detection of low-level mild indications at single-photon sensitivity and picosecond time resolution.

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The forth combination (650/ 510 nm, not shown) does not contain photons because the detection wavelength is shorter than the excitation wavelength. It should be noted here that multiplexing of lasers can also be obtained by ‘pulseinterleaved excitation’, or PIE, see Sect. 3. In that case the lasers are multiplexed pulse by pulse. The multiplexing technique shown in Fig. 16 has, however, a few advantages. The most important one is that it is free of crosstalk. 18 W. Becker The tail of the fluorescence decay excited by one laser does not show up in the decay curve excited by the next one.

Detection by two detectors at 510 and 700 nm An example is shown in Fig. 17. A leaf was excited by two multiplexed diode lasers of 405 and 650 nm wavelength. Fluorescence was detected by two detectors at 510 and 700 nm and recorded in one TCSPC module via a router. The result are three fluorescence decay curves for different combination of excitation and emission wavelength: 405/510, 405/700, and 650/700 nm. The forth combination (650/ 510 nm, not shown) does not contain photons because the detection wavelength is shorter than the excitation wavelength.

Becker Fig. 30 FLIM by wavelength multiplexing, Supercontinuum laser with AOTF, two parallel SPC-150 TCSPC modules. Left to right Excitation 500 nm emission 525 ± 50 nm, excitation 500 nm emission 620 ± 30 nm, excitation 580 nm emission 620 ± 30 nm. 256 × 256 pixels, 256 time channels, frame-by-frame multiplexing Laser pulses Laser2 blue Laser1 green Laser2 blue Laser1 green Fluorescence blue channel Fluorescence green channel Recorded Time Interval Fig. 31 Principle of laser wavelength multiplexing by pulse-interleaved excitation A similar effect can be obtained by ‘Pulse-Interleaved Excitation’, or ‘PIE’.

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